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. 2021 Mar 23;11:6608. doi: 10.1038/s41598-021-85569-9

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Defective coronary vessel organization in E12.5 Nkx2-5 mutant embryonic hearts, and the effects of hyperoxia. (A) Dorsal view of whole-mount CD31 immunostaining of ventricles showing endothelial cell plexus organized within the ventricles. CD31 staining was less prominent and extended to the apical area in Nkx2-5 mutant heart at normoxia, but was comparable at hyperoxia to the control wild-type heart (black arrowheads). (B) Quantitative CD31-positive area relative to the entire ventricular area. The number of embryos examined is indicated. The non-parametric Kruskal–Wallis ANOVA with Dunn’s test was used for analysis. (C) Representative heart sections stained with troponin-T. Thickness of compact layer is shown in white arrowheads. (D) Quantitative data showing thickness of compact layer in Nkx2-5 mutant hearts at normoxia vs. hyperoxia. The Mann–Whitney U test was used for analysis. Data are expressed as mean ± S.D.; individual data are represented as circles. *P ≤ 0.05. LV left ventricle, RV right ventricle, OFT outflow tract. The number of mice examined is indicated. ImageJ 1.52d (http://imagej.nih.gov/ij) was used for analysis.